Toxoplasma gondii (T. gondii) and Toxocara spp. are two types of parasites that can infect humans and various animals, including dogs. Police dogs and their trainers have a vital role in law enforcement, and their health and well-being are crucial for them to effectively carry out their duties. No study has yet been conducted on the prevalence of T. gondii and Toxocara spp. infections among police dogs and their trainers in Iran. The objective of this study was to determine the sero-molecular prevalence of T. gondii and Toxocara spp. infections in police dogs and their trainers in Tehran, the capital of Iran. In Tehran province, the anti-narcotics police have nearly 200 well-trained police dogs. Each dog is assigned a dedicated trainer and upon completing missions, is housed separately in a designated area. In the present study, a total of 150 samples were gathered. These included 50 blood samples from randomly selected police dogs, 50 fecal samples from the same dogs, and 50 blood samples from their trainers. The Modified Agglutination Test (MAT) was performed to detect T. gondii antibodies in dog blood samples and the ELISA system was utilized to identify anti-Toxoplasma and anti-Toxocara antibodies in the sera of the dog trainers. A specific segment of the SAG2 and ITS genes were amplified via nested-PCR in order to molecularly detect T. gondii in human blood samples and Toxocara spp. in dog fecal samples. Regarding serological findings, the prevalence of T. gondii in dog and human blood samples was 4% (2/50) and 10% (5/50), respectively. According to reports, the seroprevalence of Toxocara spp. in human blood samples was 6% (3/50). No statistically significant association was found between the prevalence of the examined parasites and variables (age, sex, and breed) in dogs, as well as the age variable in military personnel. Molecular findings showed that out of the 50 dog fecal samples and 50 human blood samples, there was no presence of Toxocara spp. and T. gondii, respectively. Understanding the prevalence of parasitic infections helps public health officials assess the risk to human and animal populations. This information can guide the development of prevention and control measures to reduce the spread of these infections. Overall, the prevalence of parasitic infections, particularly T. gondii and Toxocara spp., in police dogs and their trainers remains uncertain and necessitates further in-depth research.